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作 者:张小萍[1] 高琪[1] 王丽琴[1] 顾亚萍[1] 王伟明[1]
出 处:《中国人兽共患病杂志》1998年第4期22-25,共4页Chinese Journal of Zoonoses
基 金:世界银行/联合国开发总署/世界卫生组织热带病研究和培训特别规划处Re-entry的资助
摘 要:目的为了纯化单克隆抗体M(26-32)及其靶抗原编码基因表达的融合蛋白,使其用于疟疾快速诊断盒的研制。方法:分别用FPLC层析系统的MonoQ阴离子交换树脂柱层析方法和ImmunoAffinityIsolation柱层板技术纯化M(26-32)和其表达蛋白。结果纯化的M(26-32)有效成份为IgM,等电点在pH4.70—4.85。纯化的靶抗原基因片段表达产物为一个约130kDa的含β-半乳糖苷酶的蛋白。Western—blot分析证实纯化的M(26-32)能特异性地和其表达蛋白反应。结论纯化的表达蛋白可直接用作诊断盘研制中的质控蛋白。AIM To purify the pan - species monoclonal antiody, M26-32,and the fusion protein expressed from the target antigen gene fragment,which are going to be used in the fast immunodiagnosis kit for malaria. METH ODS:M26-32 was purified through Mono Q ion exchange column under FPLC system and the fusion protein was purifiec through an Immuno Affinity Isolation cloumn. RESULTS The effective composition of purified M(26-32) was IgM and its isoelectric point ranged from PH4. 70 to 4. 85. The purified fusion protein was a 130 kDa protein containing β-galactosidase. The purified M26-32,could specifically react with the purified fusion protein in Western -blot experiment. CONCLUSION:The purified fusion protein,as a control protein,can be directly applied in the development of a fast immunodiagnosis kit for malaria based on M(26-32).
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